Skip to main content
SpringerLink
Log in

Development of plant regeneration and transformation protocols for the desiccation-sensitive weeping lovegrass Eragrostis curvula

  • Cell Biology and Morphogenesis
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

A tissue culture protocol, suitable for transformation, was established for the pasture grass Eragrostis curvula. Callus was generated in the dark from leaf and seed tissues on a medium comprising MS salts supplemented with 2 mg/l 2,4-D, 0.01 mg/l BAP and 2% sucrose. Plant regeneration occurred via organogenesis on the same medium with 6% and 3% sucrose for shoot and root formation, respectively. Optimal regeneration (50 plantlets per callus) occurred when light of 45 μmol/m2 per s was used. The yeast Saccharomyces cerevisiae Hsp12 gene was cloned into the Sac1 of the pCAMBIAUbeeQ vector and callus was transformed by biolistic bombardment. Best transformation (30%) occurred when the target tissue was bombarded twice at a distance of 70 mm using a bombardment pressure of 9,100 kPa. Although successful transformation and transcription of the Hsp12 gene occurred, no Hsp12 protein was found present in tissue extracts of transformed grass.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1a–d
Fig. 2
Fig. 3

Similar content being viewed by others

Abbreviations

BAP:

6-Benzylaminopurine

2.4-D:

2,4-Dichlorophenoxyacetic acid

GUS:

β-Glucuronidase

HSP:

Heat shock protein

MS:

Murashige Skoog

PCR:

Polymerase chain reaction

TDZ:

Thidiazuron

X-Gluc:

1-Bromo-2-chloro-3-indoyl-β-glucuronic acid

References

  • Chen W, Lennox S, Palmer K, Thomson JA (1998) Transformation of Digitaria sanguinalis: a model system for testing maize streak virus resistance in Poaceae. Euphytica 104:25–31

    Google Scholar 

  • Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159

    Article  CAS  PubMed  Google Scholar 

  • Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15

    Google Scholar 

  • Echenique V, Polci P, Mroginski L (1996) Plant regeneration in weeping lovegrass, (Eragrostis curvula) through inflorescence culture. Plant Cell Tissue Organ Cult 46:123–130

    Google Scholar 

  • Farrant JM (2000) A comparison of mechanisms of desiccation tolerance among three angiosperm resurrection plant species. Plant Ecol 151:29–39

    Google Scholar 

  • Gaff DF (1989) Responses of desiccation tolerant “resurrection” plants to water stress. In: Kreeb KH, Richter H, Hinckley TM (eds) Structural and function responses to environmental stress: water shortage. XIV International Botanical Congress, Berlin, pp 225–268

    Google Scholar 

  • Klein TM, Wolf ED, Wu R, Sanford JC (1987) High velocity microprojectiles for delivering nucleic acid into living cells. Nature 327:70–73

    Google Scholar 

  • Mtwisha L, Brandt W, McCready S, Lindsey GG (1998) Hsp12 is LEA-like protein in Saccharomyces cerevisiae. Plant Mol Biol 37:513–521

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497

    CAS  Google Scholar 

  • Roberts CS, Rajagopal S, Smith LA, Nguyen TA, Yang W, Nugroho S, Ravi KS, Cao M-L, Mijayachandra K, Patell V, Harcourt RL, Dransfield L, Desamero N, Slamet I, Keese P, Kilian A, Jefferson RA (1998) A comprehensive set of modular vectors for advanced manipulations and efficient transformation of plants by both Agrobacterium and direct DNA uptake methods. PCAMBIA vector release manual, version 3.05

  • Sales K, Brandt W, Rumbak E, Lindsey GG (2000) The LEA-like protein in Saccharomyces cerevisiae has a plasma membrane location and protects membranes against desiccation and ethanol-induced stress. Biochim Biophys Acta 1463:267–278

    Google Scholar 

  • Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbour Laboratory Press, Cold Spring Harbour, N.Y.

    Google Scholar 

  • Vander Willigen C, Pammenter NW, Mundree S, Farrant JM (2001) Some physiological comparisons between the resurrection grass, Eragrostis nindensis, and the related desiccation sensitive species, E. curvula. Plant Growth Regul 35:121–129

    Google Scholar 

Download references

Acknowledgements

The authors thank Dr S. G. Mundree for donation of the pCAMBIAUbeeQ vector. The work was supported by a grant to Jill Farrant awarded by the National Research Foundation and the DST Innovation fund.

Author information

Authors and Affiliations

  1. Department of Molecular and Cellular Biology, University of Cape Town, Private Bag, Rondebosch, 7700, South Africa

    Sandile Ncanana, Wolf Brandt, George Lindsey & Jill Farrant

Authors
  1. Sandile Ncanana
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wolf Brandt
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. George Lindsey
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jill Farrant
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jill Farrant.

Additional information

Communicated by A. Altman

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ncanana, S., Brandt, W., Lindsey, G. et al. Development of plant regeneration and transformation protocols for the desiccation-sensitive weeping lovegrass Eragrostis curvula. Plant Cell Rep 24, 335–340 (2005). https://doi.org/10.1007/s00299-005-0940-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-005-0940-1

Keywords

Search

Navigation